CN104012753A - Active microorganism functional feed additive for livestock and preparation method and application thereof - Google Patents
Active microorganism functional feed additive for livestock and preparation method and application thereof Download PDFInfo
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- CN104012753A CN104012753A CN201310719955.8A CN201310719955A CN104012753A CN 104012753 A CN104012753 A CN 104012753A CN 201310719955 A CN201310719955 A CN 201310719955A CN 104012753 A CN104012753 A CN 104012753A
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- 244000005700 microbiome Species 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000003674 animal food additive Substances 0.000 title abstract description 5
- 240000008371 Bacillus subtilis Species 0.000 claims abstract description 59
- 229940075615 Bacillus subtilis Drugs 0.000 claims abstract description 55
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 55
- 238000000855 fermentation Methods 0.000 claims abstract description 45
- 230000004151 fermentation Effects 0.000 claims abstract description 45
- 239000000843 powder Substances 0.000 claims abstract description 37
- 239000000654 additive Substances 0.000 claims abstract description 29
- 230000000996 additive Effects 0.000 claims abstract description 29
- GUBGYTABKSRVRQ-UUNJERMWSA-N Lactose Natural products O([C@@H]1[C@H](O)[C@H](O)[C@H](O)O[C@@H]1CO)[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1 GUBGYTABKSRVRQ-UUNJERMWSA-N 0.000 claims abstract description 17
- 239000008101 lactose Substances 0.000 claims abstract description 17
- GUBGYTABKSRVRQ-XLOQQCSPSA-N lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims abstract description 17
- 241000194031 Enterococcus faecium Species 0.000 claims abstract description 16
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- 229940035034 maltodextrin Drugs 0.000 claims abstract description 9
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- BWKOZPVPARTQIV-UHFFFAOYSA-N azanium;hydron;2-hydroxypropane-1,2,3-tricarboxylate Chemical compound [NH4+].OC(=O)CC(O)(C(O)=O)CC([O-])=O BWKOZPVPARTQIV-UHFFFAOYSA-N 0.000 claims description 6
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Abstract
The invention discloses an active microorganism functional feed additive for livestock. A preparation method of the additive comprises the following steps: producing a dry active powder agent containing enterococcus faecium WEI-10 and bacillus subtilis WEI-2 by a method of submerged fermentation of liquid, subsequently mixing the agent with yeast powder, fructo-oligosaccharide, sorbitol, lactose and maltodextrin in an appropriate ratio to obtain granules, or mixing the agent with yeast powder, fructo-oligosaccharide, sorbitol, lactose and white carbon black in an appropriate ratio to obtain a powder agent, weighing and packaging to obtain the microorganism functional feed additive product. The functional feed additive adopts the strain with a definite source, is excellent in property, good in production property and high in stability, has remarkable effects, is suitable for livestock, is helpful in improving the production performance and the anti-disease capability of livestock, and can regulate the micro-ecological balance of intestinal tracts of livestock and improve the quality of products such as meat and eggs.
Description
Technical field
The present invention relates to formula, preparation method and the application of active microorganism functional feedstuff additive, belong to biology feed additive production technical field.
Background introduction
Active microorganism functional feedstuff additive utilizes useful active microorganism to improve animal health level, production performance, there are the potentiality of alternative feeding antibiotic, can ensure the safety of feed, become the novel fodder additive ensuring food safety, become the focus of people's research and development at present.
Current commercially available additive for microbe feedstuff product is very different, the source of contained bacterial classification, phylogenetic feature, probiotic properties in most products, security, effect characteristic are all unclear, and then cause the effect of product to be had a greatly reduced quality, the inexpensive product of low-quality floods market, have a strong impact on understanding and the confidence of consumer for additive for microbe feedstuff, thereby limited the promotion and application of additive for microbe feedstuff in market.Trace it to its cause is that most of manufacturers do not grasp real effectively excellent species and production technology thereof, do not verify the validity of additive for microbe feedstuff formula by actual feeding experiment effect, in this case, the product of developing is difficult to produce the beneficial effect of its publicity.
Therefore, develop a kind of strain efficacy is clear and definite, formula combination is good active microorganism functional feedstuff additive for development green bio feed, guarantee food security is significant, is also in order to meet the demand of market for high-quality additive for microbe feedstuff product simultaneously.
Summary of the invention
The present invention is directed to the deficiencies in the prior art a kind of livestock and poultry active microorganism functional feedstuff additive is provided.Wherein contained active microorganism is source and clear and definite VREF (Enterococcus faecium) WEI-10CGMCC No.7746 and bacillus subtilis (Bacillus subtilis) the WEI-2CGMCC No.7744 of characteristic, WEI-10 and WEI-2 also together send the center preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms on June 19th, 2013, numbering is respectively CGMCCNo.7746 and CGMCC No.7744, preservation address is: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of Chinese Academy of Sciences postcode: 100101 phones: 86-10-64807596.All the other compositions are commercially available quality product, wherein active yeast bacterium powder, purchased from Angel Yeast Co.,Ltd, overcomes current existing microbe additive bacterial classification source with this indefinite, and the unintelligible product stability that causes of effect characteristic is poor, survival rate is low, the problem that result of use cuts in and out.
Livestock and poultry provided by the invention comprise two kinds of formulations with active microorganism functional feedstuff additive: granule formula is as follows: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, maltodextrin 70%, wherein VREF viable count>=1 × 10
10cFU/g, bacillus subtilis viable count>=1 × 10
10cFU/g, S. cervisiae viable count>=1 × 10
9cFU/g, total living bacteria count>=2 × 10 of product
10cFU/g; Wp formula is as follows: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, white carbon 70%, wherein VREF viable count>=1 × 10
10cFU/mL, bacillus subtilis viable count>=1 × 10
10cFU/mL, S. cervisiae viable count>=1 × 10
9cFU/mL, total living bacteria count>=2 × 10 of product
10cFU/mL.Wherein said manure enterococcin strain is VREF (Enterococcus faecium) WEI-10CGMCC No.7746; Described bacillus subtilis strain is bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744.
Livestock and poultry provided by the invention comprise the steps: by the preparation method of active microorganism functional feedstuff additive
(1) bacterial classification of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 and bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 is inoculated in respectively to seed culture medium and fermentation medium separately, after liquid deep layer fermenting, collect respectively thalline, make pulvis;
(2) VREF bacterium powder step (1) being made and bacillus subtilis bacterium powder proportionally mix with saccharomycete bacterium powder, FOS, sorbierite, lactose and maltodextrin; granulate at drying condition lower sheeting; by granulator fragmentation; Vibration Screen is divided into the particle of required size, after weighing, packing, produces granular pattern microbial function forage additive product.
(3) VREF bacterium powder step (1) being made and bacillus subtilis bacterium powder proportionally mix with saccharomycete bacterium powder, FOS, sorbierite, lactose and white carbon, after weighing, packing, produce powder-type microbial function forage additive product.
Wherein, the preparation method of VREF of the present invention (Enterococcus faecium) WEI-10CGMCC No.7746 active powder is as follows: choose a ring bacterial classification from the strain inclined plane of rejuvenation, activation and be connected in the 1000mL triangular flask that 600mL seed culture medium is housed, 37 DEG C, leave standstill and cultivate 10h, viable count reaches 1 × 10
9-2 × 10
9cFU/mL, described seed culture based formulas is: in every 1000mL water, contain: peptone 20g, yeast extract 10g, glucose 20g, ammonium citrate 2g, sodium acetate trihydrate 3g, three hypophosphite monohydrate hydrogen dipotassium 1g, bitter salt 0.2g, Manganous sulfate monohydrate 0.1g, Tween 80 1g, Medium's PH Value is adjusted to 8.0, conventional sterilizing, then be connected in the 100L fermentation tank that 60L seed culture medium is housed with 1% (V/V) inoculum concentration, described seed culture medium component is consistent with shake-flask seed culture medium, sterilising conditions is 121 DEG C, 30min, fermentation condition is 37 DEG C, 50-100rpm, stuffiness, fermentation 8-10h, viable count reaches 2 × 10
9more than CFU/mL, and under similarity condition, the viable count of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 in common MRS culture medium is 7.5 × 108CFU/mL, finally be connected in the 5000L fermentation tank that 3000L fermentation medium is housed with 2% (V/V) inoculum concentration, described fermentative medium formula is to contain in every 1000mL water: peptone 20g, sucrose 20g, ammonium citrate 2g, sodium acetate trihydrate 5g, three hypophosphite monohydrate hydrogen dipotassium 2g, bitter salt 0.2g, Manganous sulfate monohydrate 0.05g, Tween 80 1g, Medium's PH Value is adjusted to 8.0, 121 DEG C of sterilizing 30min, fermentation condition is 40 DEG C, 50-100rpm, stuffiness, fermentation 8-10h, more than viable count reaches 5 × 109CFU/mL, pH is 4.0-4.5, and under similarity condition, the viable count of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 in common MRS culture medium is 9.6 × 108CFU/mL.By the nutrient solution fermenting by after high speed tube centrifuge enrichment thalline again by vacuum freeze drying processing; cryodesiccated protective agent formula is: skimmed milk power 10%; lactose 8%; Pidolidone sodium 1%; vitamin C 1%; glycerine 1%; protective agent with centrifugal after bacterium mud mix than 5: 1 according to volume mass; freeze-drying parameter is as follows: vacuum is less than 30Pa; condenser temperature is-50 DEG C to-60 DEG C, pre-freeze time 5h, drying time 40-45h; dry end, after pulverizing, obtaining viable count is 2 × 10
11the dry bacterium powder of CFU/g.
Wherein the preparation method of bacillus subtilis of the present invention (Bacillus subtilis) WEI-2CGMCC No.7744 active powder is as follows: choose a ring bacterial classification from the strain inclined plane of rejuvenation, activation and be connected in the 1000mL triangular flask that 250mL liquid seed culture medium is housed, in 35-37 DEG C, shaken cultivation 20-30h, the shake-flask seed liquid that obtains bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, viable count is 2 × 10
9cFU/mL, the state of gemma in just having started to form.The formula of described shake-flask seed culture medium is: in every 1000mL water, contain: peptone 20g, and yeast extract 10g, three hypophosphite monohydrate hydrogen dipotassium 1g, Manganous sulfate monohydrate 0.1g, pH value is 7.4, conventional sterilizing, then be equipped with in the 100L seed fermentation tank of 50L seed fermentation culture medium with the access of 1% (V/V) inoculum concentration, in 35-37 DEG C, cultivate 20-25h, speed of agitator is 250-300rpm, control suitable throughput, and add proper quantity of defoaming agent, pressure is 0.05-0.06MPa, obtain first order seed zymotic fluid, viable count is 3.2 × 109CFU/mL, the state of gemma in just having started to form, and under similarity condition, the viable count of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 in broth bouillon is 2.2 × 109CFU/mL.Described seed fermentation culture medium is consistent with the formula of shake-flask seed culture medium, and sterilising conditions is 125 DEG C, 40min, finally be equipped with in the 5000L production fermentation tank of 2500L fermentation medium with the access of 2% (V/V) inoculum concentration, in 35-37 DEG C, fermentation 25-30h, speed of agitator is 250-300rpm, control suitable throughput, and add proper quantity of defoaming agent, pressure is 0.06-0.08MPa, fermentation ends obtains the viable bacteria fermentation liquid of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, viable count is more than 5 × 109CFU/mL, gemma productive rate is 80% left and right, the formula of described fermentation medium is: in every 1000mL water, contain: peptone 20g, yeast extract 5g, glucose 10g, three hypophosphite monohydrate hydrogen dipotassium 1g, Manganous sulfate monohydrate 0.15g, bitter salt 0.3g, pH value is 7.2, 125 DEG C of sterilizing 40min, and under similarity condition, the viable count of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 in broth bouillon is 3.5 × 109CFU/mL, gemma productive rate is 62% left and right.Zymotic fluid after fermentation ends is transferred in mix and blend tank after by high speed tube centrifuge enrichment thalline, is 2: 1-3: 1 ratio adds carrier in mix and blend tank, mixes according to the mass ratio of carrier and bacterium mud.The steam pressure of spray drying device is remained on to 0.3-0.6MPa simultaneously, inlet temperature reaches 180-200 DEG C, outlet temperature reaches 110-140 DEG C, remain on-12-18Pa of tower internal pressure, then open head tank valve, start water spray, treat pressure stability, when outlet temperature remains on 80-90 DEG C, open beating pump, the mixed material of bacterium mud and carrier is squeezed into surge tank, start spraying dry, in dry run, keep the stable of tank internal pressure and out temperature, crushed after being dried obtains the dry bacterium powder of activity of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, bacterium powder viable count reaches 2 × 10
11more than CFU/g, described carrier is maltodextrin.
Use object, using method and the consumption of product of the present invention:
Product of the present invention is applicable to the routine of the livestock and poultry such as pig, chicken, duck, goose and feeds, and due to high nutritive peculiarity and multiple prebiotic effect of product, is particularly useful for in-pig, cub and poult.Mix this product thoroughly rear Direct-fed according to the ratio of 0.1-0.2 ‰ and livestock and poultry basal diet.Stress the phase, the prevention from suffering from the diseases phase, consumption doubles convalescence after being ill.
The advantage of product of the present invention and effect:
The microorganism that product of the present invention uses is the listed bacterial classification of Ministry of Agriculture's " feed addictive kind catalogue ", can use safely.The advantage of product maximum of the present invention is that bacterial classification VREF (Enterococcus faecium) the WEI-10CGMCC No.7746 using is inventor and independently separates, screens from healthy animal enteron aisle with bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, detection has the single bacterial strain of good probiotic properties and fermentation character, definite functions, effect is obvious, active high, preservation characteristic is good, and production performance is stable.Empirical tests, VREF (Enterococcus faecium) WEI-10CGMCC No.7746 can be in artificial gastro-intestinal digestion liquid normal survival 26h, can enduring high-concentration pig cholate (2%), conventional Gram-negative and positive pathogenic bacterium (Escherichia coli, salmonella and staphylococcus aureus) are all had to bacteriostasis, and antibacterial circle diameter reaches 30-40mm; Bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 exists with spore form because of it, there is the ability of the artificial gastro-intestinal digestion liquid of stronger tolerance and cholate, the more important thing is that this bacterium can secrete generation high activity amylase, protease and cellulase, this has important effect for the degraded of ANFs and the decomposition of the nondigestible material of macromolecule in promotion feed; The interpolation of S. cervisiae both can be for growth of animals or poultry provides a large number of nutrients, for example various vitamin, trace element, and amino acid, carbohydrate etc., can also provide carbon source, nitrogenous source and energy etc. for microbial growth; In product, another active component FOS is not absorbed by animal and bird intestines, also cannot be utilized by the harmful bacteria in animal and bird intestines, can only be by the lactic bacteria useful in enteron aisle and Bifidobacterium selective use, therefore the interpolation of FOS had both contributed to survival and the propagation of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 in enteron aisle, contribute to again in enteron aisle the propagation of intrinsic lactic acid bacteria itself and Bifidobacterium, thereby regulating intestinal canal microecological balance, promotes livestock birds health level.
Four kinds of active ingredients, according to best proportioning compound use, have and mutually promote, the effect of Synergistic.VREF can reduce enteron aisle pH after animal intestinal is grown surely, suppresses the growth of harmful bacteria, activates Intestinal Mucosal Immunization; Although bacillus subtilis can not be colonizated in enteron aisle, but can consume the oxygen in enteron aisle, the breeding that manufacture anaerobic environment is lactic acid bacteria creates favorable conditions, can secrete a large amount of digestive ferments and promote ANFs and the degraded of molecular nutrition material greatly in feed simultaneously, for the growth of beneficial bacterium provides nutriment, promote animal digesting and assimilating feed ingredient; Saccharomycete both can, for animal provides nutriment, can promote again the propagation of microorganism, had good symplastic growth effect with lactic acid bacteria and bacillus subtilis; FOS is a kind of good prebiotics, can optionally promote beneficial bacterium, mainly the propagation of lactic acid bacteria and Bifidobacterium, therefore four kinds of composite uses of active ingredient can produce following effect: 1) promote growth of animals or poultry, especially the growth of children livestock and poultry in age, improve efficiency of feed utilization, reduce feed cost, shorten the livestock on hand time; 2) suppress growth, the breeding of enteric pathogenic bacteria, thereby prevent and treat livestock and poultry, reduce the use of the veterinary drugs such as antibiotic; 3) improve fowl poultry immune function, the especially resistance against diseases of children livestock and poultry in age, thus reduce death rate, improve survival rate; 4) improve female livestock breed rate, thereby promote the production capacity of whole plant; 5) regulate animal and bird intestines microecological balance, prevent, extenuate dam constipation and newborn animal diarrhoea; 6) improve meat Egg Quality, improve nutrient composition content.
Brief description of the drawings
Fig. 1 is the technological process of production of a kind of livestock and poultry active microorganism functional feedstuff additive.
Detailed description of the invention
Provide embodiment below so that the present invention is specifically described; it is to be noted that following examples can not be interpreted as limiting the scope of the invention, some nonessential improvement that the person skilled in the art of this area foregoing according to the present invention is made the present invention and adjustment still belong to protection scope of the present invention.
Embodiment mono-: the preparation method of active microorganism functional feedstuff additive for livestock and poultry, comprises the steps:
(1) bacterial classification of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 and bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 is inoculated in respectively to seed culture medium and fermentation medium separately, after liquid deep layer fermenting, collect respectively thalline, make pulvis;
(2) VREF (Enterococcus faecium) WEI-10CGMCC No.7746 bacterium powder step (1) being made and bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 bacterium powder proportionally mix with saccharomycete bacterium powder, FOS, sorbierite, lactose and maltodextrin; granulate at drying condition lower sheeting; by granulator fragmentation; Vibration Screen is divided into the particle of required size, after weighing, packing, produces granular pattern microbial function forage additive product.In described granular pattern microbial function forage additive, the percentage by weight of each component is: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, maltodextrin 70%, wherein VREF viable count>=1 × 10
10cFU/g, bacillus subtilis viable count>=1 × 10
10cFU/g, S. cervisiae viable count>=1 × 10
9cFU/g, composite rear total living bacteria count>=2 × 10
10cFU/g.
(3) VREF (Enterococcus faecium) WEI-10CGMCC No.7746 bacterium powder step (1) being made and bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 bacterium powder proportionally mix with saccharomycete bacterium powder, FOS, sorbierite, lactose and white carbon, after weighing, packing, produce powder-type microbial function forage additive product.In described powder-type microbial function forage additive, the percentage by weight of each component is: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, white carbon 70%, wherein VREF viable count>=1 × 10
10cFU/mL, bacillus subtilis viable count>=1 × 10
10cFU/mL, S. cervisiae viable count>=1 × 10
9cFU/mL, composite rear total living bacteria count>=2 × 10
10cFU/mL.
Wherein, the preparation method of VREF (Enterococcus faecium) WEI-10CGMCC No.7746 active powder is as follows: choose a ring bacterial classification from the strain inclined plane of rejuvenation, activation and be connected in the 1000mL triangular flask that 600mL seed culture medium is housed, 37 DEG C, leave standstill and cultivate 10h, viable count reaches 1 × 10
9-2 × 10
9cFU/mL, described seed culture based formulas is: in every 1000mL water, contain: peptone 20g, yeast extract 10g, glucose 20g, ammonium citrate 2g, sodium acetate trihydrate 3g, three hypophosphite monohydrate hydrogen dipotassium 1g, bitter salt 0.2g, Manganous sulfate monohydrate 0.1g, Tween 80 1g, Medium's PH Value is adjusted to 8.0, conventional sterilizing; Then be connected in the 100L fermentation tank that 60L seed culture medium is housed with 1% (V/V) inoculum concentration, described seed culture medium component is consistent with shake-flask seed culture medium, sterilising conditions is 121 DEG C, 30min, fermentation condition is 37 DEG C, 50-100rpm, stuffiness, fermentation 8-10h, viable count reaches 2 × 10
9more than CFU/mL; Finally be connected in the 5000L fermentation tank that 3000L fermentation medium is housed with 2% (V/V) inoculum concentration, described fermentative medium formula is to contain in every 1000mL water: peptone 20g, sucrose 20g, ammonium citrate 2g, sodium acetate trihydrate 5g, three hypophosphite monohydrate hydrogen dipotassium 2g, bitter salt 0.2g, Manganous sulfate monohydrate 0.05g, Tween 80 1g, Medium's PH Value is adjusted to 8.0,121 DEG C of sterilizing 30min, and fermentation condition is 40 DEG C, 50-100rpm, stuffiness, fermentation 8-10h, viable count reaches 5 × 10
9more than CFU/mL, pH is 4.0-4.5.By the nutrient solution fermenting by after high speed tube centrifuge enrichment thalline again by freeze drying processing, cryodesiccated protective agent formula is: skimmed milk power 10%, lactose 8%; Pidolidone sodium 1%; vitamin C 1%, glycerine 1%, protective agent with centrifugal after bacterium mud mix than 5: 1 according to volume mass; freeze-drying parameter is as follows: vacuum is less than 30Pa; condenser temperature is-50 DEG C to-60 DEG C, pre-freeze time 5h, drying time 40-45h; dry end, after pulverizing, obtaining viable count is 2 × 10
11the dry bacterium powder of CFU/g.
Wherein the preparation method of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744 active powder is as follows: choose a ring bacterial classification from the strain inclined plane of rejuvenation, activation and be connected in the 1000mL triangular flask that 250mL liquid seed culture medium is housed, in 35-37 DEG C, shaken cultivation 20-30h, the shake-flask seed liquid that obtains bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, viable count is 2 × 10
9cFU/mL, the state of gemma in just having started to form.The formula of described shake-flask seed culture medium is: in every 1000mL water, contain: peptone 20g, and yeast extract 10g, three hypophosphite monohydrate hydrogen dipotassium 1g, Manganous sulfate monohydrate 0.1g, pH value is 7.4, conventional sterilizing; Then be equipped with in the 100L seed fermentation tank of 50L seed fermentation culture medium with the access of 1% (V/V) inoculum concentration, in 35-37 DEG C, cultivate 20-25h, speed of agitator is 250-300rpm, control suitable throughput, and add proper quantity of defoaming agent, pressure is 0.05-0.06MPa, obtain first order seed zymotic fluid, viable count is 3.2 × 10
9cFU/mL, the state of gemma in just having started to form.Described seed fermentation culture medium is consistent with the formula of shake-flask seed culture medium, and sterilising conditions is 125 DEG C, 40min; Finally be equipped with in the 5000L production fermentation tank of 2500L fermentation medium with the access of 2% (V/V) inoculum concentration, in 35-37 DEG C, fermentation 25-30h, speed of agitator is 250-300rpm, control suitable throughput, and add proper quantity of defoaming agent, pressure is 0.06-0.08MPa, fermentation ends obtains the viable bacteria fermentation liquid of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, and viable count is 5 × 10
9more than CFU/mL, gemma productive rate is 80% left and right, the formula of described fermentation medium is: in every 1000mL water, contain: peptone 20g, yeast extract 5g, glucose 10g, three hypophosphite monohydrate hydrogen dipotassium 1g, Manganous sulfate monohydrate 0.15g, bitter salt 0.3g, pH value is 7.2,125 DEG C of sterilizing 40min.Zymotic fluid after fermentation ends is transferred in mix and blend tank after by high speed tube centrifuge enrichment thalline, is 2: 1-3: 1 ratio adds carrier in mix and blend tank, mixes according to the mass ratio of carrier and bacterium mud.The steam pressure of spray drying device is remained on to 0.3-0.6MPa simultaneously, inlet temperature reaches 180-200 DEG C, outlet temperature reaches 110-140 DEG C, remain on-12-18Pa of tower internal pressure, then open head tank valve, start water spray, treat pressure stability, when outlet temperature remains on 80-90 DEG C, open beating pump, the mixed material of bacterium mud and carrier is squeezed into surge tank, start spraying dry, in dry run, keep the stable of tank internal pressure and out temperature, crushed after being dried obtains the dry bacterium powder of activity of bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, bacterium powder viable count reaches 2 × 10
11more than CFU/g, described carrier is maltodextrin.
Embodiment bis-: livestock and poultry are the application in livestock and poultry cultivation with active microorganism functional feedstuff additive
Product of the present invention has carried out a large amount of livestock and poultry feeding experiments.Broiler feeding result of the test shows, broiler chicken 1 age in days from being born is taken product of the present invention continuously, continues to and delivers for sale, can significantly improve broiler chicken average daily gain, reduces feedstuff-meat ratio; Can improve IgG in serum of broilers, IgA content, promote periphery blood T lymphocyte breeder reaction; Index and spleen index and bursa of farbricius index obviously increase; Product of the present invention can also significantly improve lactic acid bacterium number in Intestine of Broiler, reduces Escherichia coli bacterium number; After doubling, the using dosage of product of the present invention has prevention effect for manual-induced broiler colibacillosis.Laying hen feeding experiment result shows, laying hen takes product 30-60 days of the present invention after entering the peak of laying eggs continuously, can significantly improve performance in layers, comprise that the indexs such as average egg weight, average day egg production, laying rate, rate of broken eggs and feedstuff-egg ratio are all than the remarkable improvement of control group.Product of the present invention can effectively reduce the death rate of young fowl (such as meat chick and egg chick etc.).
Product of the present invention is especially obvious for the health-care efficacy of weanling pig, and before and after wean, using dosage doubles, and can obviously extenuate diarrhoea or growth retardation phenomenon because producing; Normally feed child care piglet 30 days, piglet average daily gain be can improve, feedstuff-meat ratio, the death rate and the incidence of disease reduced, after testing, the specific immunity of piglet and salivary lysozyme be obviously increase of contrast all, the lactic acid bacteria of ileum, colon and caecum and Bifidobacterium bacterium are counted showed increased, and Escherichia coli quantity declines.In-pig feeding experiment result shows, product of the present invention can effectively be extenuated sow in gravidic constipation phenomenon, increases a childbirth number, improves farrowing rate.Growing and fattening pigs feeding experiment result shows, product of the present invention can obviously improve the production performance of growing-finishing pig, shortens marketing time, improves cultivation efficiency.
Table 1 has been listed the product of the present invention drawing by experiment best recommendation amount in different phase animal and fowl fodder in the actual process of feeding.
The best recommendation amount of this product of table 1 in different phase animal and fowl fodder
Claims (7)
1. a livestock and poultry active microorganism functional feedstuff additive, is characterized in that:
A. granule formula consists of the following composition: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, maltodextrin 70%, wherein VREF viable count>=1 × 10
10cFU/g, bacillus subtilis viable count>=1 × 10
10cFU/g, S. cervisiae viable count>=1 × 10
9cFU/g, total viable count>=2 × 10 of product
10cFU/g.
B. wp formula consists of the following composition: VREF 5%, bacillus subtilis 5%, S. cervisiae 5%, FOS 10%, sorbierite 2%, lactose 3%, white carbon 70%, wherein VREF viable count>=1 × 10
10cFU/mL, bacillus subtilis viable count>=1 × 10
10cFU/mL, S. cervisiae viable count>=1 × 10
9cFU/mL, total viable count>=2 × 10 of product
10cFU/mL.
2. feed addictive according to claim 1, is characterized in that: described manure enterococcin strain is VREF (Enterococcus faecium) WEI-10CGMCC No.7746, raw material bacterium powder viable count>=2 × 10
11cFU/g; Described bacillus subtilis strain is bacillus subtilis (Bacillus subtilis) WEI-2CGMCC No.7744, raw material bacterium powder viable count>=2 × 10
11cFU/g.
3. the preparation method of a livestock and poultry use active microorganism functional feedstuff additive; it is characterized in that: the VREF bacterium powder preparing through fermenting and producing and bacillus subtilis bacterium powder are proportionally mixed with saccharomycete bacterium powder, FOS, sorbierite, lactose and maltodextrin; granulate at drying condition lower sheeting; by granulator fragmentation; Vibration Screen is divided into the particle of required size, after weighing, packing, produces granular pattern microbial function forage additive product.
4. preparation method according to claim 3, it is characterized in that: the VREF bacterium powder preparing through fermenting and producing and bacillus subtilis bacterium powder are proportionally mixed with saccharomycete bacterium powder, FOS, sorbierite, lactose and white carbon, after weighing, packing, produce powder-type microbial function forage additive product.
5. preparation method according to claim 3, it is characterized in that: the production stage of described VREF (Enterococcus faecium) WEI-10CGMCC No.7746 active powder is as follows: from rejuvenation, on the strain inclined plane of activation, choosing a ring bacterial classification is connected in the 1000mL triangular flask that 600mL seed culture medium is housed, 37 DEG C, leave standstill and cultivate 10h, described seed culture based formulas is: in every 1000mL water, contain: peptone 20g, yeast extract 10g, glucose 20g, ammonium citrate 2g, sodium acetate trihydrate 3g, three hypophosphite monohydrate hydrogen dipotassium 1g, bitter salt 0.2g, Manganous sulfate monohydrate 0.1g, Tween 80 1g, Medium's PH Value is adjusted to 8.0, conventional sterilizing, then be connected in the 100L fermentation tank that 60L seed culture medium is housed with 1% (V/V) inoculum concentration, described seed culture medium component is consistent with shake-flask seed culture medium, and sterilising conditions is 121 DEG C, 30min, fermentation condition is 37 DEG C, 50-100rpm, stuffiness, fermentation 8-10h, finally be connected in the 5000L fermentation tank that 3000L fermentation medium is housed with 2% (V/V) inoculum concentration, described fermentative medium formula is to contain in every 1000mL water: peptone 20g, sucrose 20g, ammonium citrate 2g, sodium acetate trihydrate 5g, three hypophosphite monohydrate hydrogen dipotassium 2g, bitter salt 0.2g, Manganous sulfate monohydrate 0.05g, Tween 80 1g, Medium's PH Value is adjusted to 8.0,121 DEG C of sterilizing 30min, fermentation condition is 40 DEG C, 50-100rpm, stuffiness, fermentation 8-10h.By the nutrient solution fermenting by after high speed tube centrifuge enrichment thalline again by vacuum freeze drying processing; cryodesiccated protective agent formula is: skimmed milk power 10%, lactose 8%, Pidolidone sodium 1%; vitamin C 1%; glycerine 1%, vacuum is less than 30Pa, and condenser temperature is-50 DEG C to-60 DEG C; pre-freeze time 5h; drying time 40-45h,, obtain viable count be 2 × 10
11the dry bacterium powder of CFU/g.
6. a livestock and poultry active microorganism functional feedstuff additive, is characterized in that: the application of described feed addictive in livestock and poultry cultivation whole process.
7. application according to claim 6, is characterized in that: described feed addictive is mixed complete feed according to 0.1-0.2 ‰.
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| CN104982659A (en) * | 2015-06-15 | 2015-10-21 | 广东大华农动物保健品股份有限公司 | Composition containing natural plant raw materials and preparation method and application thereof |
| CN105851484A (en) * | 2016-03-24 | 2016-08-17 | 宋介珍 | Microbial feed additive, and preparation process and application thereof |
| CN106615976A (en) * | 2017-01-09 | 2017-05-10 | 北京大伟嘉生物技术股份有限公司 | Compound premix for laying hens at laying stage used for formulating low-protein-content diet and application thereof |
| CN107173522A (en) * | 2016-03-09 | 2017-09-19 | 北京大伟嘉生物技术股份有限公司 | A kind of fermented feed containing probiotics and its preparation method and application |
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| CN104770576A (en) * | 2015-04-22 | 2015-07-15 | 张奎昌 | Preparation method for inclusion compound of probiotics-beta-cyclodextrin or derivative thereof |
| CN104982659A (en) * | 2015-06-15 | 2015-10-21 | 广东大华农动物保健品股份有限公司 | Composition containing natural plant raw materials and preparation method and application thereof |
| CN107173522A (en) * | 2016-03-09 | 2017-09-19 | 北京大伟嘉生物技术股份有限公司 | A kind of fermented feed containing probiotics and its preparation method and application |
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| CN106615976A (en) * | 2017-01-09 | 2017-05-10 | 北京大伟嘉生物技术股份有限公司 | Compound premix for laying hens at laying stage used for formulating low-protein-content diet and application thereof |
| CN108338258A (en) * | 2018-05-16 | 2018-07-31 | 广西青又青生物肥业有限公司 | A kind of farrowing sow Synbiotic feed additive and preparation method and application |
| CN109645217A (en) * | 2018-12-29 | 2019-04-19 | 广西万寿谷投资集团股份有限公司 | A kind of zinc-rich probiotic and preparation method thereof improving free-ranging chicken immune function |
| CN110959752A (en) * | 2019-12-18 | 2020-04-07 | 湖北华扬科技发展有限公司 | High-stability microecological compound preparation and preparation method and application thereof |
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